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Related Concept Videos

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

21.5K
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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Super-resolution Fluorescence Microscopy01:37

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Related Experiment Video

Updated: Feb 27, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

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Lens-based fluorescence nanoscopy.

Christian Eggeling1, Katrin I Willig1, Steffen J Sahl1

  • 1Department of NanoBiophotonics,Max Planck Institute for Biophysical Chemistry,37070 Göttingen,Germany.

Quarterly Reviews of Biophysics
|May 23, 2015
PubMed
Summary
This summary is machine-generated.

Super-resolution microscopy, or nanoscopy, overcomes the diffraction limit of light, enabling visualization of finer cellular details. This technology offers new possibilities for detailed biophysical and biomedical research on living cells.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Traditional fluorescence microscopy is limited by light diffraction, restricting spatial resolution to ~200 nm.
  • This resolution limit hinders detailed visualization of subcellular structures and molecular interactions.
  • Overcoming this limit is crucial for advancing cell biology and biomedical research.

Purpose of the Study:

  • To review the principles and modalities of current fluorescence nanoscopy techniques.
  • To discuss the potential applications of nanoscopy in biophysical and cellular experiments.
  • To highlight the impact of super-resolution microscopy on life sciences research.

Main Methods:

  • Describes various fluorescence nanoscopy techniques that surpass the diffraction limit.
  • Explains how nanoscopy variants transiently control fluorophore emission states (e.g., 'on'/'off') for feature separation.
  • Discusses the use of photoswitchable fluorophores and standard labels in different nanoscopy approaches.

Main Results:

  • Nanoscopy achieves molecular spatial resolution, overcoming previous limitations.
  • Different nanoscopy variants offer distinct advantages and disadvantages regarding fluorophore requirements and applicability.
  • Techniques allow for dynamic, multi-color, and 3D imaging of biological samples.

Conclusions:

  • Fluorescence nanoscopy represents a significant advancement for studying the living cell.
  • It enables research at an unprecedented level of detail, addressing long-standing scientific questions.
  • Lens-based fluorescence nanoscopy is expected to drive future innovations in life sciences.